摘要
目的:建立浙贝母超高效液相色谱-四极杆-飞行时间串联质谱(UPLC-Q-TOF-MS/MS)指纹图谱,并确定其抗炎质量标志物。方法:色谱柱为Eclipse Plus C18,流动相为甲醇-0.1%甲酸水溶液(梯度洗脱),流速为0.4 mL/min,柱温为30℃,进样量为2μL;采用电喷雾离子源,以正、负离子模式检测,在质荷比(m/z)50~1200范围内进行扫描。采用《中药色谱指纹图谱相似度评价系统》(2004A版)建立10批不同产地浙贝母的UPLC-Q-TOF-MS/MS指纹图谱。以小鼠耳肿胀度和血清中丙二醛(MDA)、一氧化氮(NO)水平为抗炎指标,采用双变量相关分析法和灰色关联度分析法对浙贝母指纹图谱中各共有峰相对峰面积与抗炎指标的相关性进行分析,筛选质量标志物并进行结构鉴定。结果:在正、负离子模式下,10批浙贝母药材分别各有共有峰26、10个。综合双变量相关分析和灰色关联度分析结果,共发现与抗炎作用相关的质量标志物9个,经鉴定分别为贝母素乙、贝母素甲、环巴胺、胡萝卜苷、PolyphyllinⅤ、苦鬼臼毒素、植物甾醇、Ent-kaur-15-en-17-ol、Ent-17-norkauran-16-one。结论:所建立的UPLC-Q-TOF-MS/MS指纹图谱可用于评价浙贝母药材的质量;贝母素乙、贝母素甲、环巴胺等可能是浙贝母具有抗炎作用的质量标志物。
OBJECTIVE:To establish UPLC-Q-TOF-MS/MS fingerprint of Fritillaria thunbergii,and to define its antiinflammatory quality markers.METHODS:The determination was performed on Eclipse Plus C18 column with mobile phase consisted of methanol-0.1%formic acid solution(gradient elution)at the flow rate of 0.4 mL/min.The column temperature was set at 30℃,and injection volume was 2μL.The electrospray ion source was used to scan in the range of m/z 50-1200 with positive and negative ion detection mode.UPLC-Q-TOF-MS/MS fingerprints of 10 batches of F.thunbergii from different habitats were established by using the Similarity Evaluation System of TCM Chromatographic Fingerprints(2004 A edition).With ear swelling degree,the serum levels of MDA and NO as anti-inflammatory indexes,the correlation between the relative area of common peaks in fingerprint and the anti-inflammatory indexes was analyzed by using bivariate correlation analysis and grey correlation analysis,and the quality markers were screened and identified.RESULTS:In positive and negative ion mode,10 batches of F.thunbergii had 26 peaks and 10 peaks.Based on bivariate correlation analysis and grey correlation analysis,nine quality markers related to anti-inflammatory effect were found,which were identified as peiminine,peimine,cyclobalamine,daucidin,polyphyllinⅤ,bitumen podophyllotoxin,phytosterol,ent-kaur-15-en-17-ol,ent-17-norkauran-16-one.CONCLUSIONS:Established UPLC-QTOF-MS/MS fingerprint can be used to evaluate the quality of F.thunbergii;peiminine,peimine and cyclobalamine and so on may be the quality markers of anti-inflammatory effect of F.thunbergii.
作者
程斌
周爱珍
彭昕
王昕蕾
王娟
CHENG Bin;ZHOU Aizhen;PENG Xin;WANG Xinlei;WANG Juan(School of TCM,Zhejiang Pharmaceutical College,Zhejiang Ningbo 315100,China;Ningbo Research Institute of Zhejiang University,Zhejiang Ningbo 315000,China;College of Pharmacy,Harbin University of Commerce,Harbin 150076,China)
出处
《中国药房》
CAS
北大核心
2020年第17期2129-2135,共7页
China Pharmacy
基金
浙江省基础公益研究计划项目(No.LGN19H280002)
浙江省食品药品监督管理局2018年度科技计划项目(No.2018018)
宁波市科技创新2025重大专项(No.2019B10008)。
